DE4002825C2 - Drug combination of a squalene synthetase inhibitor and another serum cholesterol lowering agent - Google Patents

Description

The invention relates to a combination of an inhibitor of squalene synthetase and a drug containing the serum Cholesterol other than by inhibiting squalene Synthetase decreases, as well as to the use of this Arz neoplastic combination in a process for lowering the Serum cholesterol level and / or for prevention or Be Treatment of arteriosclerosis.

There are several different classes of connections that Have serum cholesterol lowering properties. Some These compounds are inhibitors of the enzyme HMG CoA-Re Ductase, which plays an important role in the production of Cholesterol, such as mevastatin (US Pat. No. 3,983,140), Lo vastatin, which is also called mevinolin (US Pat. No. 4,231,938), Pravastatin (US-A-4,346,227) and velostatin, also Synvi Nolin is called (US-A-4,448,784 and 4,450,171).

Other compounds that lower the serum cholesterol level This can also be done according to a completely different mechanism than do the HMG CoA reductase inhibitors. For example  The serum cholesterol level can also be changed by the application bile acid scavengers such as cholestyramine, colestipol, DEAE-Sephadex and poly (diallylmethylamine) derivatives (US Pat. 4,759,923 and 4,027,009), or by using Antihy perlipoproteinemics such as probucol and gemfilbrozil which is obviously the mirror of the Lipopro low density lipoproteins (LDL) in the Serum decrease and / or LDL in high density lipoproteins Convert (HDL).

The aforementioned US-A-4,759,923 discloses poly (di allylmethylamine) derivatives which are bile acid scavengers, that can be used in conjunction with drugs, which changes the serum cholesterol level by others Reduce mechanisms as by trapping, such as clofibrate, Nicotinic acid, probucol, neomycin, p-aminosalicylic acid or Mevinolin, which is also called Lovastatin.

Squalene synthetase is a microsomal enzyme that affects the re Ductive dimerization of two molecules of farnesyl pyrophosphate phat (FPP) in the presence of nicotinamide adenine dinucleotide Phosphate (reduced form = NADPH) to form squalene catalyzes (C.D. Poulter and H. C. Rilling in "Biosynthesis of Isoprenoid Compounds ", Vol. I, Chapter 8, pp. 413-441, J. Wiley and Sons, 1981 with the bibliography cited therein point). This enzyme occurs in the first significant stage of the recent cholesterol biosynthetic pathway. The selek tive inhibition of this stage should be the unhindered fort important routes to isopentenyl-tRNA, ubiquinone and Dolichol allow. It could be shown that the Squalene synthetase together with HMG-CoA reductase Down-regulated receptor-mediated LDL uptake (J.R. Faust, J.L. Goldstein and M.S. Brown, Proc. Nat. Acad. Sci. USA, Vol. 76 (1979), pp. 5018-5022). This does the Proposal credible that inhibition of squalene synthetase leads to an upregulation of LDL receptor levels, as has been shown for HMG-CoA reductase, and that  this finally for treatment and prevention of Hypercho lesterinemia and arteriosclerosis should be useful.

One way to inhibitors of squalene synthetase is in the Creation of analogs of the substrate FPP. From the literature shows that the pyrophosphate unit for binding to the enzyme is essential. However, such pyrophosphates are as components of medicines due to their chemical and enzymatic lability with respect to allylic C-O cleavage as well as their sensitivity to metabolism unsuitable for phosphatases.

P. Ortiz de Montellano and co-workers. describe in J. Med. Chem., 20 (1977), pp. 243-249, the preparation of a series of substituted terpenoid pyrophosphates (Table A). The Authors showed that these compounds have competitive inhibi the enzyme squalene synthetase. These substances be still sit the unstable allylic pyrophosphate unit from FPP.

Table A

Corey and Volante, J. Am. Chem. Soc. 98: 1291-1293 (1976) established the FPP analog A and the presqualenic pyrophosphate (PSQ-PP) analogues B as inhibitors of squalene biosynthesis (presqualenic pyrophosphate is an intermediate in the conversion from FPP in squalene). These inhibitors have methylene groups in place of the allylic acid moiety of FPP and PSQ-PP, but still have the chemically and enzymatically unstable pyrophosphate binding.

Poulter and co-workers prepared the cyclopropane C (RM Sandifer et al., J. Am. Chem. Soc., Vol. 104 (1982) p. 7376-7378) which, in the presence of inorganic pyrophosphate, is an inhibitor of an intermediate analogue of the enzyme squalene synthetase represents.

Altman and co-workers, A. Bertolino and co-workers, Biochim. Bio Phys. Acta Vol. 530 (1978) p. 17-23 reported that ferns sylamin and related derivatives D hibate squalene synthetase, but there is evidence that this inhibition is not specific, and presumably with membrane rupture connected is.

CD Poulter et al., J. Org. Chem., 51 (1986), p. 4768 produced compound E as part of a synthetic procedure, but did not provide any biological data for it.

CD Poulter and KE Stremler, JACS, Vol. 109 (1987), p. 5542 describe the synthesis and biological evaluation of compounds of structure F. These compounds were evaluated as alternative substrates for avian liver farnesyl diphosphate and lemon peel cyclase.

RW McClard and CD Poulter et al., JACS, Vol. 109 (1987), p. 5544 report that the phosphinyl phosphonates G and H are competitive inhibitors of 1'-4 condensation between isopentenyl diphosphate and geranyl diphosphate prepared by the Bird liver farnesyl diphosphate synthetase is catalyzed. The phosphinyl phosphonates G and H have ki values of 19 μM and 71 μM, respectively. The authors also reported the possible isolation of farnesyl-phosphinylphosphonate I and geranyl-phosphinylphosphonate J from the enzymatic reaction of G with geranyl pyrophosphate and dimethyl allyl pyrophosphate, respectively. Structures I and J were attributed to these compounds because of their relative thin-film mobilities. The authors suggested that I could be a potential inhibitor of squalene synthetase.

TL Capson revealed in a dissertation of the Dept. of Medicine Dicinal Chemistry of the University of Utah, June 1987, Abstract, Inhaltsverzeichnung, p. 16, 17, 40-43, 48-51, to the summary, cyclopropanes of the formula

as inhibitors of intermediate analogues of squalene syn thetase.

SA Biller et al., Journal of Medicinal Chemistry, Vol. 31 (1988), pp. 1869-1871 disclose that isoprenoid (phosphinylmethyl) phosphonates (PMPs) inhibit the suqalene synthetase. These phosphonates have the structures

The subject of the present invention is a drug combination for use in lowering serum cholesterol sterol levels and in inhibiting the formation of in the treatment of atherosclerosis. The Medicine consists of an inhibitor of the enzyme squalene synthetase and another drug (here as the other, the Serum cholesterol lowering agent is called), the lowers the serum cholesterol level and / or the cholesterol inhibited rin biosynthesis, after another Me chanism as that of inhibiting the production of En zyms squalene synthetase or the enzyme 3-hydroxy-3-methyl glutaryl-coenzyme A (HMG CoA) reductase. Examples of such Other drugs are bile acid scavengers or antihy perlipoproteinemic agents that prevent the formation of LDL inhibit or convert LDL to HDL.  

The squalene synthetase inhibitor is in a Gewichtsver ratio to the other drug in the range of about 0.001: 1 to about 1000: 1 and preferably from about 0.05: 1 to about 100: 1 used.

The subject matter of the invention is furthermore the said drug substance combination for use in lowering the Se level of rum cholesterol and in the inhibition of digestion treatment of arteriosclerosis. The Administration of the drug combination is systemic, for example, orally or parenterally, over a longer period Period.

The combination of the squalene synthetase inhibitor with the other serum cholesterol lowering drug creates a surprising and unique way to inhibition or Treatment of elevated cholesterol levels and / or Ar teriosclerosis by producing additional anticholesterol nemic effects compared to those in use achieved the individual components of the combination alone can be. In addition, the combined drug the invention, the compounds with different effects mechanism for effective treatment of Cholesterol-causing disorders of various Etiology can be used.

The squalene synthetase inhibitors suitable for use in the medicaments of the invention include, but are not limited to, the compounds described by Biller et al., Supra, including isoprenoids (phosphinyl methyl) phosphonates, for example those of the formula

including the tri-acids, triesters and tripotassium and Trisodium salts thereof, as well as other suqalen synthetase inhi Bitoren, which are described in EP-A-324 421.

Other squalene synthetase inhibitors approved for use in Medicaments of the invention include the ter penoid pyrophosphates produced by P. Ortiz de Montellano and Coll., In J. Med. Chem., Vol. 20 (1977), pp. 243-249, the Farnesyl diphosphate analog A and presqualen pyrophosphate (PSQ-PP) analogues described by Corey and Volante, J. Am. Chem. Soc. Vol. 98 (1976), pp. 1291-1293, the phosphinyl phosphonates, that of R.W. McClard et al., J.A.C.S., Vol. 109 (1987), P. 5544 and the cyclopropanes described by T.L. Capson in the Dissertation of the Dept. Med. Chem. U. of Utah, Abstract, In table of contents, p. 16, 17, 40-43, 48-51, abstract,  and other known squalene synthesis tase inhibitors.

The other, the serum cholesterol-lowering drug, other than by inhibiting the enzyme HGM CoA reduct tase or the enzyme squalene synthetase acts and to Ver in the drug combination of the invention is, without being limited to it - an antihyperli be a poproteinemischer drug such as probucol and gemfibro zil or a related compound according to US-A-3,674,836, where Probucol and gemfibrozil are preferred. Next suitable bile acid scavengers, such as cholestyramine, colestipol and DEAE-Sephadex (Secholex®, Polidexide®), as well as Clofibrate, Li postable (Rhone-Poulenc), Eisai E-5050 (an N-substituted Ethanolamine derivative), imanixil (HOE-402), tetrahydrolipsta tin (THL), Istigmastanyl phosphorylcholine (SPC, Roche), Ami nocyclodextrin (Tanabe Seiyoku), Ajinomoto AJ-814 (an Azu len derivative), melinamide (Sumitomo), Sandoz 58-035, American Cyanamid CL-277,082 and CL-283,546 (disubstituted urine substance derivatives), nicotinic acid, neomycin, p-aminosalicyl acid, aspirin, poly (diallylmethylamine) derivatives, such as those described in U.S. Pat US Pat. No. 4,759,923, quaternary amine poly (diallyl dimethyl ammonium chloride) and ionenes as described in US-A-4,027,009 described, as well as others the serum cholesterol level lowering agents that reduce cholesterol through a mechanism other than the inhibition of the enzymes HGM CoA- Reduce reductase or squalene synthetase.

Preferred combinations include one of Biller's and Employee, a.a.O. described isoprenoid (phosphinylmethyl) - phosphonates with probucol or gemfibrozil.

On the disclosure of the aforementioned patents and Pa Patent applications are hereby incorporated by reference.

The drug combination of the invention may be for treatment of mammals, such as monkeys, dogs, cats, rats and humans,  be used. For this purpose it can be in the usual dosage forms such as tablets, capsules, juices or injections Preparations are formulated. These dosage forms contain also the necessary carriers, excipients, lubricants medium, buffer, antibacterial substances, fillers (like mannitol), antioxidants, (ascorbic acid or sodium bi sulfite) or similar excipients. Oral dosage forms are however, parenteral forms are also suitable.

The dose to be administered is carefully depending on the age, weight and condition of the patient as well as in the outward direction look at the route of administration, the dosage form, the turned diet and set the desired result.

With oral administration can be a satisfactory result be achieved when the squalene synthetase inhibitor in dosages in an amount ranging from about 10 to about 2000 mg, preferably from about 25 to 200 mg in Combination with the other, the serum cholesterol lowering Active ingredient is used, this in the usual Quantities used as described in the Physician's Desk Reference (PDR) (medical manual) for each such drug are given, for example in an amount in the range of about 2 mg to 7500 mg, preferably from about 2 mg to 4000 mg. The squalene synthetase inhibitor and the other, The serum cholesterol-lowering agent can thereby together in the same oral dosage form or in taken separately from separate oral dosage forms become.

A preferred oral dosage form, such as tablets or Capsules, contains the squalene synthetase inhibitor in one Amount of about 10 to 500 mg, preferably from about 25 to 200 mg and the other, the serum cholesterol lowering Active ingredient in an amount of about 2 to 3000 mg, preferably about 2 to 2000 mg.  

The drug combination can be found in the above described dosage forms either once or in on divided doses one to four times a day. Conveniently, the treatment is a low dose the drug combination started and stepped up as needed siege.

It can be made tablets of different sizes, for example, with a total weight of about 2 to 2000 mg containing one or both active ingredients in the above contained amounts, the remainder of physiological compatible carrier or other excipients according übli pharmaceutical practice. These tablets can be notched to get to partial doses. In similar Alternatively, gelatin capsules can be formulated.

Liquid formulations may be dissolved or suspended of an active substance or a combination of active substances in a conventional, for pharmaceutical administration acceptable liquid carrier can be prepared so that the desired dose in the amount of 1 to 4 teaspoons ready is provided. Such dosage forms may be given to the patient be given one to four times a day.

In another embodiment, the active ingredients for finer control of the dosing plan separated into individual doses simultaneously or in close succession be given off times. There by a controlled The blood glucose level is established and maintained will be the same result by the simultaneous Presence of the two substances achieved. The respective effect Materials may be administered individually in separate dosage forms be formulated as described above.

Specified combinations of a squalene synthetase Inhibitor and another, the serum cholesterol lowering  Active ingredient are more favorable and preferred, especially in form of tablets or capsules for oral administration.

For the formulation of the drugs, the active ingredients in the amounts described above according to usual pharmazeu practice with a physiologically acceptable carrier, Excipients, binders, preservatives, stabilizers, Flavoring and optionally other excipients Unit dose forms associated.

Examples of excipients for tablet production are:
Binders such as gum tragacanth, acacia, maize starch or gelatin, excipients such as dicalcium phosphate or cellulose, disintegrants such as corn starch, potato starch or alginic acid, lubricants such as stearic acid or magnesium stearate, sweeteners such as sucrose, aspartame, lactose or saccharin, gums, like orange, peppermint, olive oil or cherry flavor. When the dosage unit form is a capsule, it may contain, in addition to the excipients mentioned above, a liquid carrier such as a fatty acid oil. Various other substances may also be present as coatings or for otherwise modifying the physical form of the dosage unit. For example, tablets or capsules may be coated with shellac, sugar or both. A juice may contain the active ingredient, water, alcohol or a similar compatible carrier, glycerol solubilizer, sucrose for sweetening, methyl and propylparaben as a preservative, a dye and a flavoring such as cherry or orange flavor.

Some of the active ingredients described above form be known, pharmaceutically acceptable salts, such as alkali metal or other common basic salts, or acid additions salts. The mention of the basic substances therefore concludes the usual known salts, the parent compounds are essentially equivalent.  

The formulations described above can be administered for a longer period, d. H. so long, like the elevated serum cholesterol level or the arterio sclerosis persists or symptoms persist. formulations with delayed drug delivery, such amounts twice weekly, weekly or monthly also be used. One at least one to two-week intake is to achieve a success required.

The examples illustrate the invention

example 1

A squalene synthetase inhibitor formulation in the form of tablets is prepared from the following ingredients:

component Weight, mg (E, E) - [[Hydroxy (4,8,12-trimethyl-3,7,11-tridecatrienyl) phosphinyl] methyl] phosphonic acid tripotassium salt (squalene synthetase inhibitor according to Biller et al., Supra) 100 corn starch 50 gelatin 7.5 Avicel (microcrystalline cellulose) 25 magnesium stearate 2.5 185

The tablets are made from sufficient stocks Mixing the tripotassium salt and cornstarch with one prepared aqueous solution of gelatin. The mixture is then dried and ground to a fine powder the avicel and magnesium stearate are added with granulation mixed. The mixture becomes tablet in a tablet press th pressed with a content of 100 mg of active ingredient.  

Probucol tablets containing 250 mg of probucol made by conventional methods. They contain the following additives according to PDR 1988: maize starch, ethylcel cellulose, glycerol, hydroxypropylcellulose, hydroxypropylme ethyl cellulose 2910, iron oxide, lactose, magnesium stearate, microcrystalline cellulose, polysorbate 80, talc and titanium oxide.

The squalene synthetase inhibitor and the probucol tablets can be used as a combination to lower serum cholesterol Mirror and / or for the treatment of arteriosclerosis verwen be. In addition, the squalene synthetase inhibitor and probucol tablets ground to powder and put together in a single capsule can be used.

Example 2

Tablets containing the following components:

component Weight, mg (E, E) - [difluoro [hydroxy- (4,8,12-trimethyl-1,3,7,11-tridecatrienyl) -phosphinyl] -methyl] -phosphonic acid tripotassium salt (squalene synthetase inhibitor according to Biller et al Employee, above) 100 Avicel 112.5 lactose 113 corn starch 17.5 stearic acid 7 350

are made from sufficient stocks by bringing together of the squalene synthetase inhibitor, the avicel and a Part of the stearic acid produced. The lumps are ge grind and through a sieve with a mesh size of 8 mm guided. Then the lactose, the cornstarch and the rest  the stearic acid admixed. The mixture is dissolved in a Ta Blettenpresse to capsule-shaped tablets with a mass of 350 mg compressed. The tablets are being divided into Slotted halves.

Capsules containing gemfibrozil 300 mg are added customary processes. They contain the following Additives according to PDR 1988: polysorbate 80 NF, starch NF and Silica gel.

The squalene synthetase inhibitor tablets and the gemfibro Zil capsules can be given as a combination. Alterna The squalene synthetase inhibitor tablets may also be active ground to a powder and added in a single capsule together with the gemfibrozil to lower the serum cholesterol rin level and / or for the treatment of arteriosclerosis be used.

Example 3

A tablet preparation of the following composition is prepared as described in Example 1:

component Weight mg (E, E) - [difluoro [hydroxy (4,8,12-trimethyl-3,7,11-tridecatetraenyl) -phosphinyl] -methyl] -phosphonic acid tripotassium salt (squalene synthetase inhibitor according to Biller et al. , supra) 100 corn starch 50 gelatin 7.5 Avicel (microcrystalline cellulose) 25 magnesium stearate 2.5 185

The above-obtained squalene synthetase inhibitor Tablets can be used in combination with clofibrate capsules which are 500 mg clofibrate and inactive stock parts such as dye and gelatin according to PDR 1988. The squalene synthetase inhibitor and clofibrate may be present in separate dosage forms or together in a single Capsule for lowering elevated serum cholesterol levels or used for the treatment of arteriosclerosis.

Examples 4 to 6

The Squalene Synthetase Inhibitor Tablets of Examples 1, 2 or 3 may be used in combination with a cholestyramine resin 4 g Cholestyramine, Acacia, Citron acid, a dye, a flavoring, polysorbate 80, propylene glycol alginate and sucrose according to PDR 1988 contains, used. The squalene synthetase inhibitor and the cholestyramine, may be in separate dosage forms or together in a single capsule to lower the Se rum-cholesterol level or for the treatment of arterio sclerosis are used.

Examples 7 to 10

Squalene synthetase inhibitor tablets manufactured according to the Examples 1, 2 and 3, in combination with nicotine acid, colestipol, dextrothyroxine or others, the serum Cholesterol-lowering drugs to lower the Cholesterol level or for the treatment of arteriosclerosis be used.

It is clear that all of Biller and co-workers, supra. and in EP-A-324 421 discloses squalene synthetase inhibitors in Combination with any of the ones mentioned here, the serum Cholesterol-lowering agents can be used NEN.

Claims (8)

A drug combination comprising an inhibitor of Enzyme's squalene synthetase and another remedy substance that reduces serum cholesterol and / or Inhibits cholesterol biosynthesis, after one mechanism other than by inhibiting production the enzyme HMG CoA reductase or the enzyme squalene synthetase. 2. Combination according to claim 1, wherein the other remedy an antihyperlipoproteinemic agent or is an antihyperlipemic agent. 3. Combination according to claim 2, wherein the antihyperlipopro teinemic agent or antihyperlipemic agent probucol, gemfibrozil, clofibrate, dextrothyroxine or the sodium salt thereof, colestipol or the hydro chloride thereof, cholestyramine, nicotinic acid, neomycin, p- Aminosalicylic acid or aspirin. 4. The combination of claim 1, wherein the drug is a Bile acid scavenger is. 5. Combination according to claim 4, wherein the bile acid Ab cholestyramine, colestipol, DEAE-Sephadex, a Poly (diallylmethylamine) derivative, an ion or quater nary amine-poly (diallyldimethylammonium) chloride is. 6. A combination according to claim 1 or 3, wherein the inhibitor of En zyms squalene synthetase the formula
having. 7. A combination according to claim 1, wherein the inhibitor of En zyms squalene synthetase in a weight ratio to the other drug ranges from about 0.001: 1 to about 1000: 1 is present. 8. Use of a drug combination according to any one of claims 1 to 7 to lower elevated serum cholesterol sterol levels or to prevent their formation or for the treatment of arteriosclerosis.